CZ283320B6 - Deoxyspergualin trihydrochloride in beta crystalline form and process for preparing thereof - Google Patents

Abstract

The solution relates to deoxyspergualin trihydrochloride in crystalline beta form. This substance suppresses the body's immune response. The process for its production consists in dissolving deoxyspergualin trihydrochloride in aqueous ethanol at an ethanol concentration of 85 to 99.5% by volume, allowing the solution to stand for 2 to 22 days, and separating the beta form crystals. ŕ

Description

Deoxyspergualine trihydrochloride in crystalline beta form and process for its preparation

Technical field

The present invention relates to deoxyspergualine trihydrochloride in crystalline beta form, the chemical name of which is N- [4- (3-aminopropyl) aminobutyl] -2- (7-guanidinoheptanamide) -2-hydroxyethanamitrihydrochloride, and a process for its preparation. The compound has an antitumor effect and is able to suppress the immunological response of the organism.

BACKGROUND OF THE INVENTION

Deoxyspergualin can be represented by Formula I

H ₂ NCNH (CH ₂ ) ₆ CONHCHCONH (CH ₂ ) 4 NH (CH ₂ ) ₃ NH ₂ (I).

II

NH

OH

This substance is suitable for clinical use because of its very good antitumor activity and its ability to suppress the immunological response. Usually this is prepared in the form of the trihydrochloride.

The production of this hydrochloride is generally carried out according to the teachings of U.S. Pat. Nos. 4,518,532 and 4,523,015, a synthetic process utilizing the condensation of 7-guanidine heptanamide with glyoxylyl spermidine under dehydration using an acid catalyst.

In this way a powdered hydrochloride is obtained which has some undesirable properties, such as high temperature instability and hygroscopicity. According to the invention, it is possible to obtain crystalline hydrochloride which no longer has such pronounced undesirable properties, is more stable at elevated temperatures and is not so hygroscopic.

It has been shown that the hydrochloride can be obtained in crystalline form and that the crystals have a significantly lower hygroscopicity, are more stable at a higher temperature, and are easier to handle than the powdered or lyophilized product usually produced and used.

During the application, powdered deoxyspergualine hydrochloride refers to a powder obtained from an aqueous suspension of this hydrochloride by drying under reduced pressure or by freeze-drying. A syrup obtained from an aqueous suspension is a syrup-like residue obtained by evaporating an aqueous solution of said hydrochloride under reduced pressure. The water content of the syrup is typically 30% or less.

Crystalline deoxyspergualin hydrochloride is poorly hygroscopic and relatively stable at higher temperatures, crystals having a or β form.

Crystals in a form can be prepared as follows:

Deoxyspergualine hydrochloride can be obtained in various ways, one of which is described, for example, in Example 4 of U.S. Pat. No. 4,518,532.

When the powdered deoxyspergualin hydrochloride is allowed to stand in a low humidity environment, for example at a relative humidity of 52% or less at a suitable temperature, the powder crystallizes into deoxyspergualin hydrochloride crystals in form a.

The crystallization temperature is not limited to a certain temperature range, provided that it does not exceed the decomposition temperature of the material, typically a range of 0 to 30 ° C is used. The time required is 2 to 20 days, usually 10 days, depending on the amount of powder and moisture.

- 1 GB 283320 B6

The crystals in the α form can also be obtained from deoxyspergualin hydrochloride syrup. This is done by concentrating the aqueous solution of deoxyspergualin hydrochloride to a syrup, which is then allowed to stand at a low relative humidity, preferably 52% or less, to form crystals in the form of a. may be shortened by mixing the syrup during crystallization or by adding a suitable amount of crystals in the form of a.

The most suitable method for producing crystals in mold a is by adding a small amount of preformed mold crystals to a syrup having a water content of 20% or less and mixing the syrup for several hours at 30% or less relative humidity until solidification. Placing in a desiccator with silica gel terminates crystallization.

The physicochemical properties of deoxyspergualin hydrochloride in crystalline form a will be described below.

1. X-ray diffraction

An X-ray diffraction device (Shimadzu XD-610) equipped with a copper tube, a graphite monochrometer and a scintillation counter was used for detection, the results of the procedure are shown in Fig. 1 and summarized in Table 1.

Further, FIG. 1 shows the diffraction pattern for the crystals of the form α, the abscissa angle is plotted in degrees and the intensity in Kcps on the ordinate axis.

Table 1

Diffraction of crystals in the form d (10 ^'10 m) I / l d (10' ¹⁰ m) I / li

11.78 0.46 4.04 0.76 10.64 0.53 3.90 0.91 7.49 0.34 3.78 0.76 5.03 0.36 3.50 1.00 4.72 0.51 3.39 0.95 4.39 0.84 3.12 0.64

X-ray source: copper tube, 40 kV, 30 mA, λ = 1.54051 filter: graphite monochrometer.

2. Thermal analysis

The results of differential thermal analysis (DTA) and thermogravimetry (TG) obtained using a Rigaku Denki (Model TAS-100) are shown for mold crystals and are shown in Figure 2, the abscissa axis shows the temperature in degrees Celsius and the ordinate axis weight in mg or electron volts (gV).

3. Hygroscopicity

To evaluate the hygroscopicity of the powder and crystals in the mold and the change in weight of each sample after storage at different relative humidity for 24 hours was recorded. The results are shown in Table 2 below.

-2GB 283320 B6

Table 2

Weight changes at different relative humidity relative __________ weight increase in%

humidity (%) powder crystals of form a 14 1.5 31 5.9 -0.8 43 9.1 -0.4 52 12.6 -0.2 64 17.9 17.8

It can be seen from the table that the crystals of form a did not absorb water at a relative humidity of 52% or less, which means a great improvement over the powder form.

4. High temperature stability

In order to evaluate the stability of the crystalline form and at higher temperature compared to the powder form, samples were stored for 6 days at 50 ° C and then analyzed by high pressure liquid chromatography to determine the amount of decomposed substance. The results are shown in Table 3.

Table 3

Comparison of stability at higher temperature powder of mold crystal and amount (%)

100.0 amount after storage amount (%) = ---------------------- x 100.

initial quantity

As can be seen from Table 3, the crystals have a mold and considerably improved high temperature stability.

The crystalline form of β deoxyspergualin hydrochloride can be obtained as described below. The compound as such may be obtained by a variety of methods, one of which is described, for example, in U.S. Patent No. 4,518,532.

SUMMARY OF THE INVENTION

The present invention provides deoxyspergualine trihydrochloride in crystalline beta form, causing the following X-ray diffraction:

d (10 ^-10 meters) I / Ij d (10 ^-10 meters) I / I

15.77 0.32 4.14 0.70 10.91 0.42 3.66 1.00 4.88 0.39 3.31 0.52 4.67 0.53 3.21 0.51

The present invention also provides a process for preparing deoxyspergualine trihydrochloride in crystalline beta form; the process consists in dissolving the deoxyspergualine trihydrochloride in aqueous ethanol or contacting it with aqueous ethanol at a concentration of ethanol in a solution of 85 to 99.5% by volume, allowing the mixture to stand for 2 to 20 days with stirring, whereupon the resulting crystals of deoxyspergualine trihydrochloride in beta form separated.

The crystallization temperature is not critical provided that it does not decompose the deoxyspergualine hydrochloride, usually a temperature in the range of -20 to 50 ° C, preferably 5 ° C to room temperature is used. The crystallization time ranges from 2 to 20 days depending on the amount of hydrochloride, usually 10 days. The initial degree of crystallization proceeds faster with the use of a small amount of Form β crystals added to the crystallization mixture.

In this process, an aqueous suspension of deoxyspergualin hydrochloride may be used instead of the powder. In this case, the amount of water in the syrup should be considered so that the final ethanol concentration in the crystallization system is 85 to 99.5%, preferably 90 to 98% by volume.

The crystallization time can be reduced by stirring the mixture, then crystallization can be completed within a few days. The β-form crystals formed can be separated in a conventional manner, for example by suction filtration or by centrifugal force filtration.

The physico-chemical properties of the β-form crystals will be presented.

1. X-ray diffraction

Once again, a diffraction monitoring apparatus (Shimadzu XD-3A) equipped with a copper cathode, a graphite monochrometer and a scintillation counter was used, the results of which are shown in Figure 3 and Table 4.

Fig. 3 shows X-ray diffraction for β-type crystals by showing the angle of the diffraction in degrees and the ordinate axis the intensity in Kcps.

Table 4

X-ray diffraction for β crystals

d (10 ' ^10m ) I / Ii d ( ⁰ ' ^{, 0} m) Vij 15.77 0.32 4.14 0.70 10.91 0.42 3.66 1.00 4.88 0.39 3.31 0.52 4.67 0.53 3.21 0.52

radiation source: copper cathode, 40 kV, 30 mA, λ = 1,54051 filter: graphite monochrometer

2. Thermal analysis

The results of differential temperature analysis (DTA) and thermogravimetry (TG) obtained on a Shimadzu device (model DTG-40) are shown in Figure 4 for crystalline form β. or electron volts (gV).

3. Hygroscopicity

To assess the hygroscopicity of the powder and β-crystal, the change in weight of each sample after storage for 24 hours at different relative humidity was recorded. The results are shown in Table 5 below.

-4GB 283320 B6

Table 5

Weight changes at different relative humidity relative __________ weight increase in%

humidity (%) powder crystal form β 31 5.9 1.7 43 9.1 2.6 52 12.6 3.8 64 17.9 7.3 76 28.3 14.0

As shown in the table, the changes in crystal weight of Form β were lower at all relative humidity values. In addition, the crystals did not dissolve at a relative humidity of 64%, unlike the powdered form.

4. High temperature stability

To assess the stability of Form β crystals compared to that of the powder form, samples were stored at 50 ° C for 6 days, and analyzed by high pressure liquid chromatography to determine the fraction of decomposed material. The results are shown in Table 6.

Table 6

Comparison of stability at higher temperature powder crystal form β amount (%) 89,8 100,6 amount after storage amount (%) = ---------------------- x 100 ALIGN!

initial quantity

It can be seen from Table 6 that crystals of form β have a markedly improved thermal stability.

The following examples illustrate the invention.

Example 1

To a vial of 100 mg deoxyspergualin hydrochloride powder was added 0.5 mL of 95% (v / v) aqueous ethanol and the solution was allowed to stand at 5 ° C for 3 days. The crystals formed are filtered off under reduced pressure and dried under reduced pressure to give 55 mg of crystals in the form of β. The X-ray diffraction pattern yielded a curve typical of Form β crystals as shown in Figure 3.

Example 2

100 ml of an aqueous solution containing 20 g of deoxyspergualine hydrochloride are concentrated under reduced pressure to give 25 g of a syrupy residue. To this residue is added 75 ml of ethanol and 20 mg of Form β crystals as a seeding material. The mixture was then allowed to stand for 9 days at room temperature

-5 ° C 283320 B6 at 5 ° C. The crystals formed were collected in the same manner as in Example 1 and then dried, yielding 16 g of Form β crystals.

Example 3

500 ml of an aqueous solution containing 150 g of deoxyspergualine hydrochloride are concentrated under reduced pressure to 175 g of a syrupy residue. To this syrupy residue was added 1 liter of ethanol, and the resulting solution was stirred at 5 ° C for 2 days. The crystals formed are then collected by filtration while centrifuging and then dried under reduced pressure to give a total of 129 g of crystals in the form of β.

EXAMPLE 4 liters of an aqueous solution containing 950 g of deoxyspergualine hydrochloride are evaporated under reduced pressure to give 1120 g of a syrupy residue. To this residue was added 1.5 liters of ethanol, and the resulting suspension was transferred to a 10 liter vessel for crystallization. 6.5 liters of ethanol were added to the mixture, followed by stirring 2 g of Form β crystals, followed by stirring at 15 ° C for 4 days. The crystals were collected by centrifugation and dried under reduced pressure to give 880 g of Form β crystals.

Crystalline deoxyspergualine hydrochloride is a new substance with improved hygroscopicity and stability at higher temperature, thus providing better handling during processing into dosage forms and storage.

Claims (2)

PATENT CLAIMS 1. Deoxyspergualine trihydrochloride in crystalline beta form, causing subsequent X-ray diffraction d (10 ' ^10m ) I / I, d (10 ' ^10m ) VIi 15.77 0.32 4.14 0.70 10.91 0.42 3.66 1.00 4.88 0.39 3.31 0.52 4.67 0.53 3.21 0.51 2. A process for the production of deoxyspergualine trihydrochloride in crystalline beta form, which comprises: (a) dissolve deoxyspergualine trihydrochloride in aqueous ethanol or contact with aqueous ethanol at a ethanol concentration of 85 to 99,5% vol., b) The mixture is allowed to stand for 2 to 20 days with stirring, where appropriate c) separating the resulting deoxyspergualine trihydrochloride crystals in beta form.